Objective:To explore the differentiation of rat bone mesenchymal stem cells to dopaminergic neurons by chemically defined sequential culture media in vitro. Methods:The rat bone mesenchymal stem cells were isolated from rat marrow by density gradient centrifugation and the fourth generation was induced into dopaminergic neurons by treated with bFGF,EGF,NCM,SHH and FGF8. Dopamine associated proteins and genes in the treated cells were examined by Western blot,immumofluore-scence and RT-PCR. Results:After 2 weeks of stimulation,the cells expressed the characteristics of dopaminergic neurons. RT-PCR discovered the expression of mRNA of DAT,Nurr1,AADC,VAMT and TH. Immumofluorescence staining indicated that the ratio of TH positive neural cells was significantly increased after induction for 2 weeks(41.18±3.24) %. Conclusion:Chemically defined sequential culture media can induce differentiation of BMSCs into DA neurons in vitro,and the expression of mature neurons characteristics of the transdifferentiated cell. BMSCs might be a suitable and available source for the in vitro derivation of DA neurons and cell transplantation therapy on some central neural system diseases such as Parkinson’s disease.